The Kinase Enzyme Systems include a recombinant kinase enzyme, a substrate appropriate for the enzyme, a reaction buffer and supplemental reagents as needed. The DGKA Kinase Enzyme System contains:
- DGKA Kinase, 10μg (Human, recombinant; full-length); MW: ~82kDa
- Dilauroyl-sn-glycerol Substrate (10mM)
- 5X Reaction Buffer; DTT, 0.1M; Lipid Dilution Buffer II, 1.5ml; CaCl2 Solution (1M)
Recombinant full-length human DGKA was expressed by baculovirus in Sf9 insect cells using an N-terminal His tag. Diacylglycerol kinase alpha (DAG kinase alpha) is a member of the eukaryotic diacylglycerol kinase family. Upon cell stimulation it converts the second messenger diacylglycerol into phosphatidate, initiating the resynthesis of phosphatidylinositols and attenuating protein kinase C activity. Involved in hepatocellular carcinoma progression through regulation of the Ras/Raf/MEK/ERK pathway.
Application Note
NCBI Database Entry
The DGKA Kinase Enzyme System can be purchased with or without the ADP-Glo™ Kinase Assay reagents. Used together, the ADP-Glo™ Kinase Assay + Kinase Enzyme Systems provide a convenient method for profiling the effect of lead compounds on kinase activity. Assay advantages include broad dynamic range, ease of use and high sensitivity. Kinase Enzyme Systems are manufactured by SignalChem. Bulk quantities available upon request.
Use with ADP-Glo™ Kinase Assay
The ADP-Glo™ Kinase Assay is a luminescent kinase assay that measures ADP formed from a kinase reaction; ADP is converted into ATP, which is a substrate in a reaction catalyzed by Ultra-Glo™ Luciferase that produces light. The luminescent signal positively correlates with ADP amount and kinase activity. The assay is well suited for measuring the effects of chemical compounds on the activity of a broad range of purified kinases, making it ideal for both primary screening as well as kinase selectivity profiling. The ADP-Glo™ Kinase Assay can be used to monitor the activity of virtually any ADP-generating enzyme (e.g., kinase or ATPase) using up to 1mM ATP.